摘要
磷是植物生长必需的大量元素之一,几乎参与植物所有的生命活动。有效磷利用效率低是影响作物产量和地理分布的非生物胁迫之一。水稻作为世界上主要的粮食作物和单子叶植物的模式种,采用蛋白质组学方法研究水稻对低磷胁迫的应答机制,进而研究其耐性机理,对全面揭示水稻磷营养利用的遗传基础,发掘低磷应答相关基因、蛋白质和代谢物意义重大。
本研究在课题组以往研究基础上,选取磷效率差异的2个水稻品种IR74和IR64,在全磷和低磷条件下取1h、3h、6h、1d、3d、6d的根系和叶片进行蛋白质组学比较研究,旨在揭示水稻蛋白质响应低磷胁迫的应答及动态调控过程,主要研究结果如下：
1、不同磷效率水稻根系蛋白对低磷胁迫的差异表达
本研究利用双向电泳技术构建了磷高效水稻IR74和磷低效水稻IR64根系低磷胁迫六个时间点的差异表达蛋白谱。全磷培养各时期无显著变化,低磷培养与全磷培养间存在显著变化的两种磷效率水稻根系蛋白,磷高效水稻IR74共检测到26个,磷低效水稻IR64共检测到25个,其中20个蛋白点既在磷高效水稻也在磷低效水稻表现差异。鉴于这些蛋白对磷胁迫的响应和在两个基因型水稻中量的差异,可以认为它们是低磷诱导相关蛋白。取代表性强的31个根系蛋白进行了LC-MS质谱检测和数据库检索,27个根系蛋白成功鉴定,依据其在植物体生命活动中的主要功能归类为四大类群,一是参与转录调控的蛋白,4个；二是参与代谢的蛋白,10个；三是参与细胞生长的蛋白,3个；四是参与细胞防御的蛋白,10个。
2、不同磷效率水稻叶片蛋白对低磷胁迫的差异表达
全磷培养各时期无显著变化,低磷培养与全磷培养间存在显著变化的两种磷效率水稻叶片蛋白,磷高效水稻共检测到22个,磷低效水稻检测到19个,14个为磷高效水稻和磷低效水稻应对低磷胁迫诱导共同蛋白。取代表性变化的27个蛋白点进行LC-MS质谱检测,数据库检索有25个蛋白功能得到成功鉴定,依据在植物体生命活动中的主要功能归类为四类,一是参与光合作用的蛋白,7个；二是参与代谢的蛋白,8个；三是参与逆境反应的蛋白,7个；四是其它蛋白,3个。
As one of essential macronutrients for plant growth, phosphorus (P) is involved in almost all of plant life activities. Phosphorus deficiency in soil is one of the major abiotio stresses that limit plant growth and crop productivity worldwide. Rice is one of the world's major food crops, but also to monocotyledonous species. Proteomics approach to study the rice to phosphorus stress response mechanism, further study on tolerance mechanisms comprehensively reveal the genetic basis of use by the phosphorus nutrition of rice, is of great significance to explorephosphorus response genes, proteins and metabolites.
In this study, we choose IR74(high P-absorption efficiency) and IR64(low P-absorption efficiency) as the materials to investigate phosphorus stress resistance mechanism. The total proteins in the root and leaves are obtained respectively of IR74and IR64at low P stress after1hours,3hours,6hours,1days,3days,6days. All the proteins are separated by two-dimensional gel electrophoresis (2-DE) to reveal rice protein response to low phosphorus stress response and dynamic control process.The results obtained are summarized as follows:
1. The differentially expressed of root proteins in different P efficiencies rice uptake efficiency genotype
Differential expression profile of P-dificiency responsive protein between IR74and IR64is constructed by using2-D technology. As an object of study,we choose that in the culture of each period of full-phosphorus have no significant change, but low phosphorus culture the two phosphorus efficiency between culture and total phosphorus there are significant changes in rice root protein. It is found that26proteins of IR74and25proteins of IR64expressed differently.Twenty of them is found in both IR74and IR64expressed differentially in the different treatment time courses significantly. In view of these protein response to phosphorus stress in two rice genotypes in the amount of the difference, that they are low-phosphorus-related protein induced.31root proteins are detected by LC-MS and database searching,27of them are identified. According to their function in plants, those proteins are divided into4categories:4proteins in transcriptional regulation,10proteins in metabolism,3proteins in cell growth,10proteins in cellular defense.
2. The differentially expressed of leaf proteins in different P efficiencies rice uptake efficiency genotype
The results show that22proteins of IR74and19proteins of IR64expressed differently.14of them is found expressed differentially in both1R74and IR64.27of leaf proteins are detected by LC-MS and database searching,25of them are identified. According to their function in plants, those proteins are divided into four categories:7proteins in photosynthesis,8proteins in metabolism,7proteins in Stress responses,3proteins in other processes
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